RNA Isolation and Characterization Protocols

1. Front Matter

   Pages i-xiii

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2. Introduction to Isolating RNA

   • Donald E. Macfarlane, Christopher E. Dahle

   Pages 1-6

3. Large and Small Scale RNA Preparations from Eukaryotic Cells

   • Wolfgang Uckert, Wolfgang Walther, Ulrike Stein

   Pages 7-14

4. An Improved Rapid Method of Isolating RNA from Cultured Cells

   • David B. Batt, Gordon G. Carmichael, Zhong Liu

   Pages 15-18

5. Isolating RNA with the Cationic Surfactant, Catrimox-14

   • Christopher E. Dahle, Donald E. Macfarlane

   Pages 19-21

6. RNA Extraction from Formalin-Fixed and Paraffin-Embedded Tissues

   • Giorgio Stanta, Serena Bonin, Rosella Perin

   Pages 23-26

7. Extraction and Purification of RNA from Plant Tissue Enriched in Polysaccharides

   • Shu-Hua Cheng, Jeffrey R. Seemann

   Pages 27-32

8. Isolation of Plant Mitochondrial RNA from Green Leaves

   • Fei Ye, Wolfgang O. Abel, Ralf Reski

   Pages 33-38

9. Extraction of RNA from Fresh and Frozen Blood

   • Bimal D. M. Theophilus

   Pages 39-45

10. Isolation of Total RNA from Bacteria

    • John Heptinstall

    Pages 47-53

11. Isolation of Total RNA from Tissues or Cell Lines

    • Tapas Mukhopadhyay, Jack A. Roth

    Pages 55-59

12. Isolation of Messenger RNA

    • Sian Bryant, David L. Manning

    Pages 61-64

13. UV Spectrophotometric Analysis of Ribonucleic Acids

    • Ralph Rapley, John Heptinstall

    Pages 65-68

14. Formaldehyde Gel Electrophoresis of Total RNA

    • Sian Bryant, David L. Manning

    Pages 69-72

15. Preparation of RNA Dot-Blots

    • Rachel Hodge

    Pages 73-75

16. Nonradioactive Northern Blotting

    • Rainer Löw

    Pages 77-86

17. The Use of RNA Probes for the Analysis of Gene Expression

    • Dominique Belin

    Pages 87-102

18. Analysis of RNA by Northern Blotting Using Riboprobes

    • Rai Ajit K. Srivastava

    Pages 103-112

19. RNA Quantitative Analysis from Fixed and Paraffin-Embedded Tissues

    • Giorgio Stanta, Serena Bonin, Renè Utrera

    Pages 113-119

20. Quantitative Analysis of RNA Species by PCR and Solid-Phase Minisequencing

    • Anu Suomalainen, Ann-Christine Syvänen

    Pages 121-131

Ribonucleic acids are central to cellular and molecular processes and perform vital functions in both structural and functional roles. RNA molecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecules that deliver the amino acid building blocks to the ribosomes. Of all the RNA species, the nucleic acid intermediate, messenger RNA, is a desirable source of material to biologists, since this reflects much of, what ultimately, is translated into enzymes and proteins. In order to determine the qualitative and quantitative changes in mRNA expression, a vast number of molecular biological techniques have been developed. Key molecular methods that provide the means to initially isolate and analyze RNA molecules are the focus of this volume. In putting together this collection of protocols, we have tried to provide techniques that are most applicable and widely used. In particular, there are a number of iso- tion techniques included that have been developed, modified, or adapted to enable extraction from a variety of cell types, organisms, or subcellular organelles. Successful isolation of intact RNA is an essential starting point for any sub- quent analysis. This is why we have aimed to make this section comprehensive. The analysis of RNA is the focus of the following chapters.

• University of Hertfordshire, Hatfield, UK

  Ralph Rapley

• Tenovus Institute for Cancer Research, Cardiff, UK

  David L. Manning

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